| Summary: | During the transformation of wild-type (WT) <i>Arabidopsis thaliana</i>, a T-DNA containing <i>OLEOSIN-GFP</i> (<i>OLE1-GFP</i>) was inserted by happenstance within the <i>GBSS1</i> gene, resulting in significant reduction in amylose and increase in leaf oil content in the transgenic line (OG). The synergistic effect on oil accumulation of combining <i>gbss1</i> with the expression of <i>OLE1-GFP</i> was confirmed by transforming an independent <i>gbss1</i> mutant (GABI_914G01) with <i>OLE1-GFP.</i> The resulting <i>OLE1-GFP</i>/<i>gbss1</i> transgenic lines showed higher leaf oil content than the individual <i>OLE1-GFP/</i>WT or single <i>gbss1</i> mutant lines. Further stacking of the lipogenic factors WRINKLED1, Diacylglycerol O-Acyltransferase (DGAT1), and Cys-OLEOSIN1 (an engineered sesame OLEOSIN1) in <i>OG</i> significantly elevated its oil content in mature leaves to 2.3% of dry weight, which is 15 times higher than that in WT Arabidopsis. Inducible expression of the same lipogenic factors was shown to be an effective strategy for triacylglycerol (TAG) accumulation without incurring growth, development, and yield penalties.
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